Fig. 5
Genetic (Knock-down) and chemical (inhibitor) blockage of CD44 results in a reduction of THP-1 transmigration across HBMEC. a, b Down-regulating effect of the CRISPR Cas9 system was measured by Western blotting. β-actin was used as a loading control for each sample. Results showed a significant decrease (77 %) in the CD44/β-actin optical density ratio (***P <0.001). c Effect of Cn and HIV-1 gp41 on the BBB permeability, as evaluated by TEER. HBMEC, SC HBMEC or KD-CD44 HBMEC cultures were treated with Cn (2 × 107 CFU/ml), HIV-1 gp41 (20 μm) alone or Cn in combination with HIV-1 gp41 for 6 h. Either Cn or HIV-1 gp41 alone or joint use of them had no significant effect on the permeability of HBMEC, SC HBMEC and KD-CD44. d Effect of down-regulated CD44 expression on ZO-1 in HBMEC. e Transmigration assays were performed with HBMEC, SC HBMEC and KD-CD44, a significant suppression of transmigration was observed in the KD-CD44 group. f Bikunin inhibits Cn- and HIV-1 gp41-I90- induced THP-1 transmigration across HBMEC in a dose-dependent manner. An uninfected BMEC as a negative control was designed in the assay. Results are expressed as the mean and standard deviation of quadruplicate assays. (*P < 0.05, **P <0.01, ***P < 0.001)